Stable DNA constructs for expression of .alpha.-1 antitrypsin
AssigneeZymoGenetics, Inc.
InventorsGlenn Kawasaki, Leslie Bell
Life of this patent
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Abstract
Methods are provided for producing .alpha.-1-antitrypsin in host cells and for selecting transformed cells comprising the step of transforming the host cell with a DNA molecule comprising a gene which complements a deficiency in the host cell. The host cell is a strain having a deficiency in a function necessary for normal cell growth. The gene in the DNA molecule, such as a plasmid, which complements the deficiency serves as a selectable marker whereby the growth conditions for selection may comprise a conventional complex medium.
Claims6
A DNA construct comprising a yeast CDC4 gene which complements a deficiency in a microorganism host cell, said deficiency being in a function necessary for normal cell growth in a complex medium not containing antibiotics or heavy metals and not depleted of specific nutrients, and a DNA sequence coding for alpha-1-antitrypsin, wherein said gene can function as a selectable marker in a host cell transformed with said construct to allow normal growth of said transformed cell in said medium.
A method for producing alpha-1-antitrypsin in a microorganism host cell having a deficiency in a function necessary for normal cell growth on complex media comprising the steps of:
- (a) transforming said host cell with a DNA molecule comprising a yeast CDC4 gene which complements said deficiency and a sequence coding for alpha-1-antitrypsin;
- (b) culturing the cells from step (a) under normal growth conditions in a growth medium not containing antibiotics or heavy metals and not depleted of specific nutrients, whereby said gene functions as a selectable marker for cells transformed with said DNA molecule.
A method for producing alpha-1-antitrypsin in a host cell having a deficiency in a function necessary for normal cell growth on complex media comprising the steps of:
- (a) transforming said host cell with a DNA molecule comprising a Schizosaccharomyces pombe triose phosphate isomerase gene which complements said deficiency and a sequence coding for alpha-1-antitrypsin;
- (b) culturing the cells from step (a) under normal growth conditions in a growth medium not containing antibiotics or heavy metals and not depleted of specific nutrients, whereby said gene functions as a selectable marker for cells transformed with said DNA molecule.
A DNA construct comprising a Schizosaccharomyces pombe triose phosphate isomerase gene which complements a deficiency in a microorganism host cell, said deficiency being in a function necessary for normal cell growth in a complex medium not containing antibiotics or heavy metals and not depleted of specific nutrients, and a DNA sequence coding for alpha-1-antitrypsin, wherein said gene can function as a selectable marker in a host cell transformed with said construct to allow normal growth of said transformed cell in said medium.
Dependencies are inferred from the claim text.